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Anti-Dopamine beta Hydroxylase antibody (ab43868)

Overview

  • Product nameAnti-Dopamine beta Hydroxylase antibodySee all Dopamine beta Hydroxylase primary antibodies ...
  • Description
    Rabbit polyclonal to Dopamine beta Hydroxylase
  • Tested applicationsIHC-Fr, IHC-P, IHC-R, IHC-FoFr, WB, ICC/IF more details
  • Species reactivity
    Reacts with: Mouse, Rat, Cow
  • Immunogen

    Full length native protein (purified) from adrenal medulla (Cow)

  • Positive controlRat brainstem, cerebellum and adrenal medulla.
  • General notesDilute the antibody with PBS or Tris buffer at a dilution no higher than 1/10, divide into aliquots and freeze.

Properties

Applications

Our Abpromise guarantee covers the use of ab43868 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Notes
IHC-Fr IHC-Fr: Use at an assay dependent dilution.
IHC-P IHC-P: 1/100 - 1/200. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
IHC-R IHC-R: Use at an assay dependent dilution. PubMed: 20503062
IHC-FoFr IHC-FoFr: 1/2000 - 1/4000. 1/2000 - 1/10000 (free-floating vibratome sections, ABC / DAB detection). Fixative: 4% paraformaldehyde in 0.1M phosphate buffer, pH 7.4; 500 mL over ~ 20 min. Post Fixation: 1.5 hour at 4°C in 4% paraformaldehyde in 0.1 M phosphate buffer, pH 7.4.If needed, low levels of glutaraldehyde (0.1-0.3%) may be used in conjunction with paraformaldehyde.
WB WB: Use a concentration of 1 - 2 µg/ml. Detects a band of approximately 72 kDa (predicted molecular weight: 68 kDa).
ICC/IF ICC/IF: 1/400 - 1/800. 1/400 - 1/800, in PBS/0.3% Triton X-100.

Target

  • FunctionConversion of dopamine to noradrenaline.
  • PathwayCatecholamine biosynthesis; (R)-noradrenaline biosynthesis; (R)-noradrenaline from dopamine: step 1/1.
  • Involvement in diseaseDefects in DBH are the cause of dopamine beta-hydroxylase deficiency (DBH deficiency) [MIM:223360]; also known as norepinephrine deficiency or noradrenaline deficiency. This disorder is characterized by profound deficits in autonomic and cardiovascular function, but apparently only subtle signs, if any, of central nervous system dysfunction.
  • Sequence similaritiesBelongs to the copper type II ascorbate-dependent monooxygenase family.
    Contains 1 DOMON domain.
  • Cellular localizationCytoplasmic vesicle > secretory vesicle lumen. Cytoplasmic vesicle > secretory vesicle > chromaffin granule lumen and Cytoplasmic vesicle > secretory vesicle membrane. Cytoplasmic vesicle > secretory vesicle > chromaffin granule membrane.
  • Target information above from: UniProt accession P09172 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links
  • Alternative names
      DBH antibodyDBM antibodyDopamine beta hydroxylase antibody
      Dopamine beta monooxygenase antibodyDopamine beta-hydroxylase (dopamine beta-monooxygenase) antibodyDopamine beta-monooxygenase antibodyDOPO_HUMAN antibodyOTTHUMP00000022501 antibodySoluble dopamine beta-hydroxylase antibody
    see all

Anti-Dopamine beta Hydroxylase antibody images

  • Ab43868 at 1/10000 dilution staining rat tissue; positive neurons in rat brainstem, vibratome sectioned, ABC / DAB detection.
  • ICC/IF image of ab43868 stained PC12 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab43868, 5µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • Anti-Dopamine beta Hydroxylase antibody (ab43868) at 1/500 dilution + Brain: pituitary gland (Human) Whole Cell Lysate - adult normal tissue (ab29749) at 10 µg

    Secondary
    Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (HRP), pre-adsorbed (ab97080) at 1/5000 dilution
    developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 68 kDa
    Observed band size : 72 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 20 kDa,76 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 12 minutes
  • ab43868 staining Dopamine beta Hydroxylase in 12 week old murine ovary tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).

    Tissue was fixed with Modified Davidson's Fixative and a heat mediated antigen retrieval step was performed using sodium citrate. Samples were then permeabilized using 0.1% Tween in TBS, blocked with 10% serum for 1 hour at 23°C and then incubated with ab43868 at a 1/200 dilution for 12 hours at 4°C. The secondary used was a biotinylated goat anti-rabbit IgG (H+L) used at a 1/200 dilution.

    See Abreview

  • ab43868 was used to detect Dopamine beta Hydroxylase in rat brain tissue for Electron Microscopy (EM). The Tissue was Perfusion fixed with 4% PFA and the stored for less than 1 week in 1% PFA refrigerated, tissue blocked with PBS-BSA for 15 min, incubation was 1 hour with primary antibody at room temperature. A 10nm Gold-conjugated antirabbite polyclonal(1/20) was used a secondary antibody.

    See Abreview

References for Anti-Dopamine beta Hydroxylase antibody (ab43868)

This product has been referenced in:
  • Polanski W  et al. The exceptional properties of 9-methyl-beta-carboline: stimulation, protection and regeneration of dopaminergic neurons coupled with anti-inflammatory effects. J Neurochem : (2010). ICC/IF ; Mouse . Read more (PubMed: 20374418) »
  • Maison SF  et al. Mice lacking adrenergic signaling have normal cochlear responses and normal resistance to acoustic injury but enhanced susceptibility to middle-ear infection. J Assoc Res Otolaryngol 11:449-61 (2010). IHC-R ; Mouse . Read more (PubMed: 20503062) »

See all 2 Publications for this product

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Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (12 Week old Mouse Ovary)
Specification 12 Week old Mouse Ovary
Fixative Modified Davidson's Fixative
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Sodium Citrate
Permeabilization Yes - 0.1% Tween in TBS
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 23°C
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Submitted Jan 31 2012

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"